U.S. patent number 4,879,272 [Application Number 06/784,640] was granted by the patent office on 1989-11-07 for method and composition for preventing the adsorption of a medicine.
This patent grant is currently assigned to Chugai Seiyaku Kabushiki Kaisha. Invention is credited to Tsutomu Kawaguchi, Naoto Shimoda.
United States Patent |
4,879,272 |
Shimoda , et al. |
* November 7, 1989 |
**Please see images for:
( Certificate of Correction ) ** |
Method and composition for preventing the adsorption of a
medicine
Abstract
A method of preventing erythropoietin in an aqueous solution
from being adsorbed on the inner surface of the wall of a container
by incorporating in the aqueous solution one or more additives, and
an erythropoietin composition so formulated as to avoid the
adsorption of erythropoietin onto the inner surface of the wall of
a container are disclosed. The additives which are useful in this
invention include human serum albumin, bovine serum albumin,
lecithin, dextrans, ethylene oxide-propylene oxide copolymers,
hydroxypropyl cellulose, methylcellulose, polyoxyethylene
hydrogenated castor oils, polyethylene glycols and the like. A
Inventors: |
Shimoda; Naoto (Saitama,
JP), Kawaguchi; Tsutomu (Saitama, JP) |
Assignee: |
Chugai Seiyaku Kabushiki Kaisha
(Tokyo, JP)
|
[*] Notice: |
The portion of the term of this patent
subsequent to February 21, 2006 has been disclaimed. |
Family
ID: |
16589697 |
Appl.
No.: |
06/784,640 |
Filed: |
October 4, 1985 |
Foreign Application Priority Data
|
|
|
|
|
Oct 9, 1984 [JP] |
|
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59-210460 |
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Current U.S.
Class: |
514/7.7; 436/8;
514/776; 514/970; 530/380; 530/395; 514/15.2 |
Current CPC
Class: |
A61K
38/1816 (20130101); A61K 47/24 (20130101); A61K
47/42 (20130101); A61K 47/36 (20130101); A61K
47/44 (20130101); A61K 47/10 (20130101); A61K
47/38 (20130101); Y10S 514/97 (20130101); Y10T
436/10 (20150115) |
Current International
Class: |
A61K
38/18 (20060101); A61K 47/10 (20060101); A61K
47/36 (20060101); A61K 47/44 (20060101); A61K
47/24 (20060101); A61K 47/42 (20060101); A61K
47/38 (20060101); A61K 037/02 (); C07K
015/14 () |
Field of
Search: |
;424/99,101
;514/8,970,814,774,776 ;530/380,395 ;436/8 |
Other References
Horne, cited in Chem. Abstracts, vol. 102:163283h, 1985. .
Beglinger et al., cited in Chem. Abstracts vol. 99:16961u 1983.
.
Feyerhard et al., cited in Chem. Abstracts vol. 95:49281t, 1981.
.
Hara et al., cited in Biol. Abstracts vol. 71(4) Feb. 15, 1981, No.
21876. .
Konwalinka et al., cited in Biol. Abstracts vol. 78(6), Sep. 15,
1984, No 40961. .
Do et al., cited in Biol. Abstracts vol. 74(12), Dec. 15, 1982, No.
80937. .
Krystal et al., British Journal of Haemotology; vol. 58, No. 3,
Nov. 1984, 533-546. .
Beglinger et al., Digestive Diseases and Sciences, vol. 28, No. 4,
Apr. 1983. .
Horne, Thrombosis Research 37; 201-212, 1985. .
Krieglstein et al., Arzneimittel-Forschung, vol. 22; No. 9, 1972,
1538-1540. .
Mizutani et al., Journal of Pharmaceutical Sciences, vol. 67, No.
8, Aug. 1978, 1102-1105..
|
Primary Examiner: Griffin; Ronald W.
Assistant Examiner: Stone; Jacqueline M.
Attorney, Agent or Firm: Morgan & Finnegan
Claims
What is claimed is:
1. A method of preventing erythropoietin in an aqueous solution
from being adsorbed on the inner surface of the wall of a container
by incorporating in the aqueous solution one or more additives
selected from the group consisting of human serum albumin and
bovine serum albumin.
2. A method according to claim 1 wherein said one or more additives
are incorporated in amounts of 0.001-1% (w/v) of the
erythropoietin-containing aqueous solution.
3. An erythropoietin-containing, pharmaceutically-acceptable
composition wherein human serum albumin is mixed with
erythropoietin either during the preparation of said composition or
just before administration thereof.
4. An erythropoietin-containing composition according to claim 3
wherein said bovine serum albumin is incorporated in amounts of
0.001-1% (W/V) of an aqueous solution in which said composition is
dissolved.
Description
BACKGROUND OF THE INVENTION
The present invention relates to a method for preventing
erythropoietin in an aqueous solution from being adsorbed on the
inner surface of the wall of a container. The invention also
relates to an erythropoietin composition so formulated as to avoid
the adsorption of erythropoietin onto the inner surface of the wall
of a container.
Erythropoietin is a circulating glycoprotein that stimulates the
formation of red blood cells and is useful in the treatment or
diagnosis of anemia. A single dose of erythropoietin is as small as
a few micrograms and this level must be strictly observed. In other
words, erythropoietin must be administered in the accurate trace
amount in which it is formulated in a dosage form.
However, it has been observed that erythropoietin in an aqueous
solution adsorbs on the inner surface of the wall of a glass or
plastic container, and the amount that is actually administered is
significantly smaller than the amount initially formulated in a
dosage form. For example, when an aqueous solution of
erythropoietin is charged into a glass or plastic container for
transfusion, a considerable amount of erythropoietin is adsorbed on
the inner surface of the container wall and this is more likely to
occur with lower concentrations of erythropoietin than higher
concentrations. Erythropoietin which must be administered in a
trace amount has a great chance of adsorption on the inner surface
of the wall of a container and because of the smallness of the
amount in which it is initially incorporated, the loss of
erythropoietin due to adsorption is substantial, causing a
significant decrease in the dose that can be actually administered.
The necessary and accurate amount of erythropoietin could be
dissolved in aqueous solution by taking great care to avoid any
adsorption of the erythropoietin on the inner surfaces of the wall
of containers and other devices used in the preparation of aqueous
erythropoietin solutions. However, when the aqueous solution of
erythropoietin is transferred into a glass or plastic syringe for
actual administration or if it is injected into a glass or plastic
container for mixing with a transfusion solution, a substantial
portion of erythropoietin is adsorbed on the inner surface of the
wall of the syringe or container, causing serious problems for
therapeutic purposes.
Under these circumstances, the present inventors made various
studies to develop a method for preventing erythropoietin in
aqueous solution from being adsorbed on the inner surface of the
wall of a container.
SUMMARY OF THE INVENTION
In accordance with one of its aspects, the present invention
provides a method of preventing erythropoietin in aqueous solution
from being adsorbed on the inner surface of the wall of a container
by incorporating in the aqueous solution one or more additives
selected from the group consisting of human serum albumin, bovine
serum albumin, lecithin, dextrans, ethylene oxide-propylene oxide
copolymers, hydroxypropyl cellulose, methylcellulose,
polyoxyethylene hydrogenated castor oils and polyethylene glycols.
In accordance with another aspect, the invention provides an
erythropoietin-containing composition that prevents erythropoietin
from being adsorbed on the inner surface of the wall of a
container. The composition capable of achieving this object has one
or more of the additives listed above which is mixed with
erthropoietin either during the preparation of the composition or
just before its administration.
BRIEF DESCRIPTION OF THE DRAWINGS
The FIGURE is a graph showing the relationship between the
concentration of human serum albumin, which is one of the additives
specified by the present invention, and the percentage of recovery
of erythropoietin in the aqueous solutions prepared in Experiments
1 and 2.
DETAILED DESCRIPTION OF THE INVENTION
In accordance with the present invention, the erythropoietin may be
obtained by any known method; it may be extracted from the human
urine, followed by separation and purification; alternatively, it
may be produced in E. coli, yeasts or Chinese hamster ovary cells
by the genetic engineering technology, and extracted from the
culture by a variety of methods, followed by separation and
purification. The dosage of erythropoietin varies with the object
of a specific diagnosis or treatment, and formulations containing
0.1-50 .mu.g of erythropoietin per ml may be used. It should
however be noted that the present invention is not limited by the
erythropoietin content.
As will be shown more specifically by the Experiments given later
in this specification, the additive used in the present invention
for the purpose of preventing the adsorption of erythropoietin on
the inner surface of the wall of a container is at least one member
selected from the group consisting of human serum albumin, bovine
serum albumin, lecithin, dextrans, ethylene oxide-propylene oxide
copolymers, hydroxypropyl cellulose, methylcellulose,
polyoxyethylene hydrogenated castor oils and polyethylene glycols.
These substances were found to be effective among the many
substances that were checked for their ability to prevent the
adsorption of erthropoietin on the inner surfaces of the walls of
containers. Although the exact mechanism by which these substances
prevent erythropoietin from being adsorbed on the inner surfaces of
the walls of a containers is not clear, the inventors have found
that these substances are specifically selected from attaining the
purpose of preventing the adsorption of erythropoietin and that
they prevent erythropoietin's adsorption almost completely even if
they are used at low concentrations. The additive selected from the
group of the substances listed above is desirably incorporated in
an amount of at least 0.001% of an aqueous solution in which the
erythropoietin-containing composition is dissolved. On the other
hand, these substances are desirably used in sufficiently low
concentrations so that they will not exhibit pharmacological
effects when they are administered to the human body. Therefore,
the additive is desirably incorporated in an amount not exceeding
1% of the aqueous erythropoietin solution. It should however be
noted that the concentration of the additive is by no means limited
to this particular range. As mentioned earlier, the mechanism by
which the additive used in the present invention exhibits the
ability to prevent erythropoietin's adsorption is not clearly
defined, but the effect is obviously the result of its certain
action on the inner surface of the wall of the container which
contacts the erythropoietin because the same effect is obtained
even if the additive is incorporated in the aqueous erythropoietin
solution after it is put into a container. This suggests that the
minimum necessary amount of the additive in accordance with the
present invention is determined not by the content of
erythropoietin but by the area of the inner surface of the wall of
the container in which the aqueous solution of erythropoietin is
put. There is no way to specify the maximum surface area of every
container in which erythropoietin is to be put for administration
to humans, but as will be shown in Experiments 1 and 2 given later
in this specification, the purposes of the present invention can be
satisfactorily attained if the additive is incorporated in an
amount within the above stated range, i.e., 0.001-1% of an aqueous
solution in which the erythropoietin-containing composition is
dissolved.
There exist several dosage forms in which the composition of the
present invention can be formulated. Erythropoietin and the
additive in accordance with the present invention need not be
incorporated in the same composition during its preparation.
Instead, the two may be prepared as separate entities which are
mixed just before administration. Specific examples of the dosage
form of the erythropoietin-containing composition are listed
below:
(1) a form in which erythropoietin and the additive in accordance
with the present invention are incorporated in the same aqueous
solution during preparation;
(2) a form which is so designed that separate aqueous solutions of
erythropoietin and the additive in accordance with the present
invention are mixed just before administration;
(3) a form which is so designed that erythropoietin and the
additive in accordance with the present invention are incorporated
in the same free-dried powder during its preparation and that the
powder is mixed with a separately prepared vehicle for
reconstitution just before administration;
(4) a form which is so designed that separate freeze-dried powders
of erythropoietin and the additive in accordance with the present
invention are dissolved in aqueous solutions which are mixed
together just before administration; and
(5) a form which is so designed that a freeze-dried powder of
erythropoietin and an aqueous solution of the additive in
accordance with the present invention are mixed just before
administration.
These aqueous solutions and freeze-dried powders may be readily
prepared by any of the conventional techniques that are suitable
for specific types of preparations. It is within the scope of the
invention to add suitable stabilizers or buffers to aqueous
solutions, or add buffers to freeze-dried powders, or add suitable
excipients for facilitating the freeze-drying operation.
The present invention also provides a method of preventing
erythropoietin in aqueous solution from being adsorbed on the inner
surface of the wall of a container. In order to prevent
erythropoietin's adsorption, the additive that is separately
prepared in accordance with the present invention may be added to
an erythropoietin-containing transfusion or ampule solution, or
alternatively, erythropoietin may be added to a separately prepared
transfusion or ampule solution containing the additive in
accordance with the present invention. Both methods are included
within the concept of the already described invention of an
anti-adsorption composition.
The advantages of the present invention are hereunder described
with reference to the Experiments.
Experiment 1
Material:
Test samples were prepared by dissolving 0.1%, 0.02%, 0.01%, 0.005%
and 0.003% of human serum albumin in PBS solutions. As a control, a
pure PBS solution was prepared. Method:
Polyethylene test tubes were filled with 200 .mu.l of the test
samples. Ten microliters of PBS solution containing 10 .mu.g
(14,000 cpm) of erythropoietin from human urine reductively
methylated with .sup.14 C-formaldehyde were put into each of the
test tubes, which were then left to stand at room temperature. At
10 minutes, as well as 1, 2, 5 and 20 hours, 20-.mu.l portions were
collected from each sample and their radioactivities were measured
to determine the percentage of erythropoietin recovery relative to
the value for zero minutes.
Results:
The experimental results are shown in the accompanying figure,
wherein the BLANK was the pure PBS solution and HSA stands for
human serum albumin. The figure clearly shows that human serum
albumin, one of the additives specified by the present invention,
was an effective agent for the purpose of preventing the adsorption
of erythropoietin on the inner surface of the wall of a test
tube.
Experiment 2
Additives within the scope of the present invention were dissolved
in PBS solutions in the concentrations shown in the Table given
below. The samples so prepared were put into polypropylene test
tubes, and after addition of erythropoietin solutions as in
Experiment 1, the tubes were left to stand at room temperature for
2 hours. The percentages of erythropoietin recovery from the
respective samples are shown in the following table.
TABLE ______________________________________ Concentration Percent
recovery Additives % of erythropoietin
______________________________________ Lecithin 0.01 78.6 0.15 91.9
Methylcellulose 0.005 80.3 0.025 88.2 Polyethylene hydro- 0.03 76.5
genated castor oil, 0.05 92.4 POE (60) Bovine serum albumin 0.005
81.7 0.02 98.8 Dextran 40 0.001 74.6 0.20 93.0 Ethylene oxide- 0.02
73.1 propylene oxide 0.10 87.5 copolymer, F68 Polyethylene 0.05
69.2 glycol 6000 0.5 76.8 None -- 16.5
______________________________________
The above data show that the additives within the scope of the
invention were effective in preventing erythropoietin from being
adsorbed on the inner surface of the wall of a polypropylene test
tube.
The following Examples are provided to further illustrate the
present invention.
EXAMPLE 1
An aqueous solution containing 5 g of mannitol, 1 mg or
erythropoietin, 100 mg of human serum albumin, 2.154 mg of sodium
acetyltryptophanate and 1.33 mg of sodium caprylate in 100 ml was
aseptically prepared. One-milliliter portions of this solution were
charged into vials and freeze-dried, followed by hermetical sealing
of the vials.
EXAMPLE 2
Freeze-dried samples of erythropoietin were prepared as in Example
1 except that 100 mg of human serum albumin was replaced by an
equal amount of bovine serum albumin.
EXAMPLE 3
An aqueous solution containing 100 mg of dextran 40, 5 g of
sorbitol and 1 mg of erythropoietin in 100 ml was aseptically
prepared. One-milliliter portions of the solution were charged into
vials and freeze-dried, followed by hermetical sealing.
EXAMPLE 4
An aqueous solution containg 1 mg of erythropoietin, 500 mg of
polyethylene glycol 4000, 30 mg of ethylene oxide-propylene oxide
copolymer and 800 mg of sodium chloride in 100 ml of 0.05M
phosphate buffered solution (pH, 7.0) was aseptically prepared.
One-milliliter portions of the solution were charged into ampules,
which were sealed by fusing.
EXAMPLE 5
An aqueous solution containing 0.5 mg of erythropoietin, 50 mg of
lecithin and 1 g of glycine in 50 ml of 0.05M phosphate buffered
solution (pH, 7.0) was aseptically prepared. The solution was
distributed among vials in 0.5-ml portions and freeze-dried,
followed by hermetical sealing of the vials. A 0.1% aqueous
solution of polyoxyethylene hydrogenated castor oil was aseptically
prepared and 1-ml portions of the solution were charged into
ampules which were sealed by fusion so as to prepare vehicles for
reconstitution.
EXAMPLE 6
An aqueous solution containing 0.5 mg of erythropoietin, 1 g of
glycine and 1 g of sorbitol in 50 ml of 0.05M phosphate buffered
solution (pH, 7.0) was aseptically prepared. The solution was
distributed among vials in 0.5-ml portions and freeze-dried,
followed by hermetical sealing of the vials. A 0.1% aqueous
solution of methylcellulose was aseptically prepared and 1-ml
portions of the solution were charged into ampules which were
sealed by fusing so as to prepared vehicles for reconstitution.
EXAMPLE 7
An aqueous solution containing 0.5 mg of erythropoietin, 1 g of
sorbitol, 25 mg of human serum albumin and 100 mg of hydroxypropyl
cellulose in 50 ml of 0.05M phosphate buffered solution (pH, 7.0)
was aseptically prepared. The solution was distributed among vials
in 0.5-ml portions and freeze-dried, followed by hermetical sealing
of the vials.
* * * * *